title: Efficient myogenic differentiation of human adipose-derived stem cells by the transduction of engineered MyoD protein.
abstract: Human adipose-derived stem cells (hASCs) have great potential as cell sources for the treatment of muscle disorders. To provide a safe method for the myogenic differentiation of hASCs, we engineered the MyoD protein, a key transcription factor for myogenesis. The engineered MyoD (MyoD-IT) was designed to contain the TAT protein transduction domain for cell penetration and the membrane-disrupting INF7 peptide, which is an improved version of the HA2 peptide derived from influenza. MyoD-IT showed greatly improved nuclear targeting ability through an efficient endosomal escape induced by the pH-sensitive membrane disruption of the INF7 peptide. By applying MyoD-IT to a culture, hASCs were efficiently differentiated into long spindle-shaped myogenic cells expressing myosin heavy chains. Moreover, these cells differentiated by an application of MyoD-IT fused to myotubes with high efficiency through co-culturing with mouse C2C12 myoblasts. Because internalized proteins can be degraded in cells without altering the genome, the myogenic differentiation of hASCs using MyoD-IT would be a safe and clinically applicable method.
authors.affiliation: Korea Research Institute of Bioscience and Biotechnology, 125 Gwahak-ro, Yuseong-gu, Daejeon 305-806, Republic of Korea.
authors.forename: Min Sun
authors.initials: MS
authors.lastname: Sung
authors.forename: Ji-Young
authors.initials: JY
authors.lastname: Mun
authors.forename: Ohsuk
authors.initials: O
authors.lastname: Kwon
authors.forename: Ki-Sun
authors.initials: KS
authors.lastname: Kwon
authors.forename: Doo-Byoung
authors.initials: DB
authors.lastname: Oh
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000595:Amino Acid Sequence; D000818:Animals; D002454:Cell Differentiation; D002467:Cell Nucleus / Q000378:metabolism; D018920:Coculture Techniques; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D017570:MyoD Protein / Q000378:metabolism*; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014161:Transduction, Genetic
chemical_list: D017570:MyoD Protein; D010455:Peptides; D011993:Recombinant Fusion Proteins
keywords: Endosomal escape; Human adipose-derived stem cells; MyoD; Myogenic differentiation; Protein transduction domain